Purification and preliminary X-ray crystallographic studies of β-microseminoprotein from human seminal plasma
June 4th, 2009 Posted by: admin
[beta]-Microseminoprotein ([beta]-MSP) is a small cysteine-rich protein with a molecular mass of 10 kDa. It was first isolated from human seminal plasma and has subsequently been identified from several species. Comparison of the amino-acid sequences of [beta]-MSP proteins suggests that the protein is a rapidly evolving protein. The function of [beta]-MSP is poorly understood. Furthermore, no crystal structure has been reported of any [beta]-MSP; therefore, determination of the crystal structure of [beta]-MSP is the foremost task in order to understand the function of this protein completely. Here, the purification, crystallization and preliminary X-ray diffraction analysis of [beta]-MSP from human seminal plasma are described. The protein was purified using anion-exchange and size-exclusion chromatography and the purified protein was crystallized using 0.1 M ammonium sulfate, 0.1 M HEPES buffer pH 7.0 and 20%(w/v) PEG 3350. The crystals belonged to the tetragonal space group P4322 and contained three [beta]-MSP molecules in the asymmetric unit. X-ray intensity data were collected to 2.4 Å resolution.
